This application requests support for Dr. Nora Engel, a junior investigator starting up her laboratory as an independent investigator. The study of epigenetic mechanisms will lead to an understanding of how the genome functions as a developmental blueprint and how perturbations of gene expression patterns can lead to cancer. The goal of this proposal is to elucidate epigenetic mechanisms leading to allele-specific gene silencing in a cluster of imprinted genes. Imprinted genes have parental-specific monoallelic expression. This proposal focuses on the Kcnq1 domain, in which a non-coding, antisense RNA is expressed from the paternal chromosome and silences its neighboring genes. We will first exploit transgenic RNA interference technology to test whether transcription of Kcnq1ot, an imprinted antisense non-coding RNA produced from exon 11 of Kcnq1, is required to maintain the imprinting at this locus. Second, we will study the physical interactions between regulatory elements at the Kcnq1 domain through chromosome conformation capture technology, testing the hypothesis that there are allele-specific interactions. Mice with a targeted mutation in the region will be compared to the wild-type mice. A third aim is to apply a combination of bioinformatic, biochemical and in vivo approaches to identify regulatory elements in the Kcnq1 domain. This proposal will open avenues for research that will improve our understanding of imprinting dysregulation in cancer. It will also serve the immediate career goal of the applicant of establishing herself as an independent investigator in an emerging field. With the help of the excellent career development environment available, and through the expertise and collaborations she will develop, the applicant will pursue the long term goal of becoming an academic leader in cancer epigenetics.